Journal Article

Screening, purification and characterization of cellulase from cellulase producing bacteria in molasses

Roy, Narayan, Islam, Farjana

2018

Overview

Objectives This study was conducted to isolate, screening and purification of cellulase from bacteria present in sugar industry waste (molasses) and characterization by morphological and biochemical analysis. Results Based on experiments, three bacterial strains produced clear transparent zone into carboxymethyl cellulose (CMC) agar plate were identified as cellulase producing bacteria. Different culture parameters such as pH, temperature, incubation period, substrate concentration and carbon sources were optimized for enzyme production. According to the morphological and biochemical tests, the isolated strains were identified as Paenibacillus sp., Bacillus sp. and Aeromonas sp. The first strain Paenibacillus sp. showed high potentiality for maximum cellulase production (0.9 µmol ml −1  min −1 ) at pH 7.0 after 24 h of incubation at 40 °C in a medium containing 1.0% CMC. Then Paenibacillus sp. was selected for enzyme purification by ammonium sulfate precipitation, DEAE-cellulose and CM-cellulose column chromatography, respectively. In last step of purification, specific activity, recovery and purification fold were 2655 U/mg, 35.7% and 9.7, respectively. The molecular weight of the purified cellulase was found to be 67 kDa by SDS-PAGE, had an optimal pH and temperature at 7.0 and 40 °C. According to substrate specificity, the purified cellulase had high specificity on CMC substrate which indicated it to be an endo-β-1,4-glucanase.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Aeromonas / Aeromonas - enzymology / Ammonium sulfate / Bacillus / Bacillus - enzymology / Bacteria / Biochemical analysis / Biomedical and Life Sciences / Biomedicine / Carbon / Carbon sources / Carboxymethylcellulose / Cell culture / Cellulase / Cellulase - isolation & purification / Cellulase - metabolism / Cellulose / Chromatography / Column chromatography / Enzyme Stability / Enzymes / Fermentation / Gel electrophoresis / Hydrogen-Ion Concentration / Industrial wastes / Life Sciences / Medicine/Public Health / Methods / Molasses / Molasses - microbiology / Molecular weight / Morphology / Paenibacillus / Paenibacillus - enzymology / pH effects / Purification / Purification and characterization / Research Note / Screening / Sodium lauryl sulfate / Strains (organisms) / Substrate Specificity / Sugar / Sugar industry / Temperature